Traffic control of completely assembled MHC class I molecules beyond the endoplasmic reticulum

Abstract

It is generally assumed that MHC class I molecules arrive at the plasma membrane following biosynthesis, assembly and architectural editing in the endoplasmic reticulum by constitutive forward movement without requirement for specific signals (bulk flow). If this is true then all overexpressed completely assembled class I molecules should arrive at the cell surface. To study the itinerary of class I traffic beyond the endoplasmic reticulum, mammalian cells that overexpress 20 to 50-fold higher amounts of the constituent heavy and light chains were established. Thorough biochemical analyses revealed that such overexpressed molecules assemble with authentic peptides that contain the canonical class I binding anchor motif in almost 1:1 stoichiometry and impart thermal stability to the heterotrimeric complex. Despite complete assembly, however, only a fraction of the overexpressed molecules reaches the cell surface. Almost all of the overexpressed class I molecules are sialylated, thus traffic as far as the trans-Golgi or the trans-Golgi network. Overexpression of class I molecules do not seem to cause a “traffic jam” in the exocytic pathway because the kinetics of traffic of Sindbis virus structural proteins to the plasma membrane are almost identical when comparing the non-engineered and engineered cells. Thus the steady state expression of class I molecules at the cell surface is further controlled either in the Golgi apparatus or at the plasma membrane.